Protocol for IPO11 deletion and re-expression in H460 lung cancer cells using CRISPR-Cas9 and plasmid transfection

Exploring the essential role of Importin 11 (IPO11) in the nuclear translocation of its potential cargo proteins requires an efficient means of IPO11 deletion and re-expression. Here, we present a protocol for the generation of IPO11 deletion using CRISPR-Cas9 and re-expression using plasmid transfe...

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Detalles Bibliográficos
Autores principales: Zhang, Yarui, Ou, Guomin, Zhou, Zhou, Wang, Jingdong, Yang, Hongying
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10206203/
https://www.ncbi.nlm.nih.gov/pubmed/37195868
http://dx.doi.org/10.1016/j.xpro.2023.102317
Descripción
Sumario:Exploring the essential role of Importin 11 (IPO11) in the nuclear translocation of its potential cargo proteins requires an efficient means of IPO11 deletion and re-expression. Here, we present a protocol for the generation of IPO11 deletion using CRISPR-Cas9 and re-expression using plasmid transfection in H460 non-small cell lung cancer cells. We describe steps for lentiviral transduction of H460 cells, single clone selection, and expansion and validation of cell colonies. We then detail plasmid transfection and validation of transfection efficiency. For complete details on the use and execution of this protocol, please refer to Zhang et al.(1)

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